GNNQQNY: Methodology for biophysical and structural understanding of aggregation

GNNQQNY sequence offers crucial information about the formation and structure of an amyloid fibril. In this study, we demonstrate a reproducible solubilisation protocol where the reduction of pH to 2.0 resulted in the generation of GNNQQNY monomers. The subsequent ultracentrifugation step removes the residual insoluble peptide from the homogeneous solution. This procedure ensures and allows the peptides to remain monomers till their aggregation is triggered by adjusting the pH to 7.2. The aggregation kinetics analysis showed a distinct lag-phase that is concentration-dependent, indicating nucleation-dependent aggregation kinetics. Nucleation kinetics analysis suggested a critical nucleus of size ~7 monomers at physiological conditions. The formed nucleus acts as a template for further self-assembly leading to the formation of highly ordered amyloid fibrils. These findings suggest that the proposed solubilisation protocol provides the basis for understanding the kinetics and thermodynamics of amyloid nucleation and elongation in GNNQQNY sequences. This procedure can also be used for solubilising such small amyloidogenic sequences for their biophysical studies.

Electrodeposition of Cu-Ag Alloy Films at n-Si(001) and Polycrystalline Ru Substrates

Electrodeposition of Cu-Ag films from acidic sulfate bath was conducted at n-Si(001) and polycrystalline Ru substrates. Significant nucleation overpotential of 0.4 V is observed with the Cu-Ag bath at n-Si(001) substrate, whereas the electrodeposition of Cu-Ag at Ru substrate is influenced by Ru oxides at the surface. Incomplete coverage of Si substrate by Cu-Ag deposit was observed from the deposition systems without Ag(I), or with 0.1 mM Ag(I), comparing with the compact Cu-Ag film obtained with the deposition bath containing 0.01 mM Ag(I). Layered and faceted Cu-Ag deposit was observed at small Cu deposition overpotential with the Ru substrate. Phase composition of the Cu-Ag deposits at n-Si(001) substrate from electrolyte with various Ag(I) concentrations is examined by XRD. Limited solubility of Ag (0.4 at.%) was observed in fcc-Cu until phase separation occurs. The classical model for nucleation kinetics in electrodeposition was used to examine the potentiostatic transients of the Cu-Ag electrodeposition at n-Si(001) substrate.

Heterogeneous Crystal Nucleation from the Melt in Polyethylene Oxide Droplets on Graphite: Kinetics and Microscopic Structure

It is well known that the crystallization of liquids often initiates at interfaces to foreign solid surfaces. In this study, using polarized light optical microscopy, atomic force microscopy (AFM), and wide-angle X-ray scattering (WAXS), we investigate the effect of substrate–material interactions on nucleation in an ensemble of polyethylene oxide (PEO) droplets on graphite and on amorphous polystyrene (PS). The optical microscopy measurements during cooling with a constant rate explicitly evidenced that the graphite substrate enhances the nucleation kinetics, as crystallization occurred at approximately an 11 °C higher temperature than on PS due to changes in the interactions at the solid interface. This observation allowed us to conclude that graphite induces heterogeneous nucleation in PEO. By employing the classical nucleation theory for analysis of the data with reference to the amorphous PS substrate, the obtained results indicated that the crystal nuclei with contact angles in the range of 100–117° were formed at the graphite interface. Furthermore, we show that heterogeneous nucleation led to a preferred orientation of PEO crystals on graphite, whereas PEO crystals on PS had isotropic orientation. The difference in crystal orientations on the two substrates was also confirmed with AFM, which showed only edge-on lamellae in PEO droplets on graphite compared to unoriented lamellae on PS.